Journal: Scientific reports

Article Title: Immunoglobulin superfamily member 3 is required for the vagal neural crest cell migration and enteric neuronal network organization.

doi: 10.1038/s41598-023-44093-8

Figure Lengend Snippet: Figure 2. Both IGSF3 mRNA and protein are expressed in the neural crest cells. (A–F) Single cell RNA sequencing analysis at E8.5 and E9.5 represented as t-SNE plots reveals the neural crest (NC) and neural tube (NT) cell clusters defined by Sox10 (B,E) and Sox2 (C,F) expression, respectively. The subclusters are assigned according to the annotations from the original analysis of the source data23. Igsf3 is expression is restricted to the neural crest cells at E8.5 (A). At E9.5, Igsf3 is expressed predominantly by the delaminating and migrating neural crest which is identified by Sox10 with some expression in the neural tube cells (F). (G,H) Serial sections of E9.5 embryo were stained using the anti-IGSF3 and anti-SOX10 antibodies (red color). IGSF3 expression (G) is detected in the Sox10-positive (H) neural crest cells as indicated by arrows pointing to the neural crest cells that are newly migrating out of the dorsal neural tube on both sides. Scale bars: (G,H) 50 µm.

Article Snippet: The primary antibodies include polyclonal rabbit IGSF3 (1:750 HPA036305, Sigma-Aldrich), polyclonal mouse SOX10 (1:500 SC-365692, Santa Cruz).

Techniques: RNA Sequencing, Expressing, Staining

Journal: Scientific reports

Article Title: Immunoglobulin superfamily member 3 is required for the vagal neural crest cell migration and enteric neuronal network organization.

doi: 10.1038/s41598-023-44093-8

Figure Lengend Snippet: Figure 3. Single-cell RNAseq analysis shows that Igsf3 is the only IGSF member that is expressed by the neural crest cells. (A) Large panel in the left upper corner displays the t-SNE embedding showing the cell clusters of E8.5 Wnt1-Cre; R26Tomatoembryo. The image is adopted from the original analysis and annotations of the source data, of which our analysis is based on23. The early neural crest comprises of three spatially distinct clusters including a Hox-negative (–) corresponding to the anterior cranial neural crest, Hoxb2- positive ( +) corresponding to the mandibular level, and HoxD3-positive ( +) corresponding to the post-otic (including cardiac and vagal) streams of the neural crest23. Single cell RNA sequence analysis reveals the expression of different Igsf members in the distinct cell clusters defined respectively by Sox10 (NC cells) and Sox2 expression (NT cells). Igsf3 is the only family member expressed solely by the neural crest cells at E8.5. The other IGSF members are expressed about equally by both cell clusters or preferentially by the neural tube cell cluster. Expression of Igsf5 and Igsf23 was not detected at E8.5. (B) Large panel in the left upper corner displays the t-SNE embedding showing the cell clusters of mouse E9.5 Wnt1Cre/R26RTomato embryo and reflects the spatiotemporal properties of neural crest (NC) development, as adopted from the original analysis of the source data23. These neural crest cell clusters are classified into the following major subpopulations such as pre-EMT, delaminating and migrating neural crest, sensory neurons, autonomic neurons, and mesenchyme23. Single cell RNA sequence analysis at E9.5 represented as t-SNE plot reveals the expression of various IgSF members in different neural tube (NT) and neural crest (NC) cell clusters defined by Sox10 and Sox2 expression, respectively. Igsf3 is expressed in the NC and its derivative cell clusters also at this stage. Igsf8 and Igsf9 are predominantly expressed in the NT population, while Igsf10 and Igsf9b are detected about equally in the NC and NT cells, and Igsf11 shows a slight preferential expression in the NC cell cluster. (C) At E10.5, the choroid plexus and NC cells specifically express Igsf3, but other Igsf members are not co-expressed in these cells.

Article Snippet: The primary antibodies include polyclonal rabbit IGSF3 (1:750 HPA036305, Sigma-Aldrich), polyclonal mouse SOX10 (1:500 SC-365692, Santa Cruz).

Techniques: Sequencing, Expressing

Journal: Annals of translational medicine

Article Title: The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway.

doi: 10.21037/atm.2020.02.14

Figure Lengend Snippet: Figure 3 IGSF3 affects HCC cell proliferation in vitro. (A,B) Western blot analyses of cyclin A, cyclin E, CDK1, and CDK2 levels in

Article Snippet: Table S4 Primary antibodies with indicated concentration for WB, IHC, IF Antibody Dilution Company Cat No. WB IHC IF IGSF3 1:400 1:400 1:50 Bioss Bs-9196R PCNA 1:250 BOSTR BM0104 cyclin A 1:250 BOSTER A00700 cyclin E 1:400 BOSTER A00543-1 CDK1 1:400 BOSTER BM0027 CDK2 1:400 BOSTER PB0562 p-IKBα 1:400 Wanleibio WL03120 IKBα 1:400 Wanleibio WL02495 IKBα 1:200 Beyotime AF1282 p-NF-κB 1:400 Wanleibio WL02169 NF-κB 1:400 Wanleibio WL01273b NF-κB 1:200 Beyotime AF1234 GAPDH 1:800 BOSTER BM3876 Ki-67 1:200 BOSTER PB0065 Lamin B 1:1,000 Bioss bs-1840R β-action 1:800 Bioss bsm-33036M Table S5 Relationship between IGSF3 expression and clinicopathologic features (n=49) Features Relative IGSF3 expression P value Low (n=23) High (n=26) Age 0.2092 ≤50 4 9 >50 19 17 Sex 0.1944 Male 22 21 Female 1 5 AFP (μg/L) 0.5276 ≤400 15 20 >400 8 6 HBV 0.7471 Positive 7 6 Negative 16 20 Tumor diameter (cm) 0.0001 ≤5 19 7 >5 4 19 TNM stage 0.0002 I–II 18 6 III–IV 5 20 Vascular invasion 0.1250 Yes 17 24 No 6 2 Lymph node metastasis 0.0045 Yes 0 8 No 23 18 Significance figures are P values for Fisher’s exact test.

Techniques: In Vitro, Western Blot

Journal: Discover oncology

Article Title: Single-cell RNA sequencing analysis reveals potential key prognostic markers in hepatocellular carcinoma.

doi: 10.1007/s12672-024-01646-1

Figure Lengend Snippet: Fig. 7 Expressions of screened genes in HCC cell lines A–B, The expression levels of BSG, CDCA8, IGSF3, PBK, and PIGZ in HCC cells were detected by RT-qPCR and western blot assays. Data are shown as means ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Primary antibodies experimented in the study were as follows: BSG (1:1000, 13,287) and PBK (1;1000, 4942) were purchased from Cell Signaling Technology (China), PIGZ (1:1000, 17,384–1-AP), CDCA8 (1:500, 12,465-1-AP), and GAPDH (1:5000, 10,494-1-AP) were obtained from Proteintech (China), IGSF3 (1:1000, ab97704) was acquired from Abcam (UK).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Annals of Translational Medicine

Article Title: The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway

doi: 10.21037/atm.2020.02.14

Figure Lengend Snippet: Primary antibodies with indicated concentration for WB, IHC, IF

Article Snippet: Primary antibodies with indicated concentration for WB, IHC, IF ( ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Company Cat No. WB IHC IF IGSF3 1:400 1:400 1:50 Bioss Bs-9196R PCNA 1:250 BOSTR BM0104 cyclin A 1:250 BOSTER A00700 cyclin E 1:400 BOSTER A00543-1 CDK1 1:400 BOSTER BM0027 CDK2 1:400 BOSTER PB0562 p-IKBα 1:400 Wanleibio WL03120 IKBα 1:400 Wanleibio WL02495 IKBα 1:200 Beyotime AF1282 p-NF-κB 1:400 Wanleibio WL02169 NF-κB 1:400 Wanleibio WL01273b NF-κB 1:200 Beyotime AF1234 GAPDH 1:800 BOSTER BM3876 Ki-67 1:200 BOSTER PB0065 Lamin B 1:1,000 Bioss bs-1840R β-action 1:800 Bioss bsm-33036M Open in a separate window caption a8 Primary antibodies with indicated concentration for WB, IHC, IF Cell proliferation, migration, and Matrigel invasion assay We used the Costar 24 well insert system (Life Technologies, corning, USA) to perform cell migration assay.

Techniques: Concentration Assay

Journal: Annals of Translational Medicine

Article Title: The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway

doi: 10.21037/atm.2020.02.14

Figure Lengend Snippet: IGSF3 affects HCC cell proliferation in vitro. (A,B) Western blot analyses of cyclin A, cyclin E, CDK1, and CDK2 levels in HCLLM3 and hepG2 cells. Bar graph shows cyclin A, cyclin D, CDK1, and CDK2 levels in HCLLM3 and hepG2 cells transfected with either shCON or shIGSF3; (C,D) FACS analysis results showing proportion of cells in each phase of the cell cycle. Bar graph shows the results of the number of cells in each phase of the cell cycle. All experiments were conducted three times. Results were indicated as mean ± SD (*, P<0.05; **, P<0.01; ***, P<0.001).

Article Snippet: Primary antibodies with indicated concentration for WB, IHC, IF ( ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Company Cat No. WB IHC IF IGSF3 1:400 1:400 1:50 Bioss Bs-9196R PCNA 1:250 BOSTR BM0104 cyclin A 1:250 BOSTER A00700 cyclin E 1:400 BOSTER A00543-1 CDK1 1:400 BOSTER BM0027 CDK2 1:400 BOSTER PB0562 p-IKBα 1:400 Wanleibio WL03120 IKBα 1:400 Wanleibio WL02495 IKBα 1:200 Beyotime AF1282 p-NF-κB 1:400 Wanleibio WL02169 NF-κB 1:400 Wanleibio WL01273b NF-κB 1:200 Beyotime AF1234 GAPDH 1:800 BOSTER BM3876 Ki-67 1:200 BOSTER PB0065 Lamin B 1:1,000 Bioss bs-1840R β-action 1:800 Bioss bsm-33036M Open in a separate window caption a8 Primary antibodies with indicated concentration for WB, IHC, IF Cell proliferation, migration, and Matrigel invasion assay We used the Costar 24 well insert system (Life Technologies, corning, USA) to perform cell migration assay.

Techniques: In Vitro, Western Blot, Transfection

Journal: Annals of Translational Medicine

Article Title: The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway

doi: 10.21037/atm.2020.02.14

Figure Lengend Snippet: IGSF3 promotes HCC cell cycle and NF-B signaling pathways in vivo. (A) Western blot analyses of cyclin A, cyclin E, CDK1, and CDK2 in liver tissue from tumor xenografts; (B) bar graph indicates data from cyclin A, cyclin D, CDK1, and CDK2 expression; (C) Western blot analyses NF-κB, p-NF-κB, IKBα and p-IKBα in liver tissue from tumor xenografts; (D) bar graph show the results of p-IKBα/IKBα and p-NF-κB/NF-κB expression. All experiments were performed three times. Results were represented as mean ± SD (*, P<0.05; **, P<0.01; ***, P<0.001).

Article Snippet: Primary antibodies with indicated concentration for WB, IHC, IF ( ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Company Cat No. WB IHC IF IGSF3 1:400 1:400 1:50 Bioss Bs-9196R PCNA 1:250 BOSTR BM0104 cyclin A 1:250 BOSTER A00700 cyclin E 1:400 BOSTER A00543-1 CDK1 1:400 BOSTER BM0027 CDK2 1:400 BOSTER PB0562 p-IKBα 1:400 Wanleibio WL03120 IKBα 1:400 Wanleibio WL02495 IKBα 1:200 Beyotime AF1282 p-NF-κB 1:400 Wanleibio WL02169 NF-κB 1:400 Wanleibio WL01273b NF-κB 1:200 Beyotime AF1234 GAPDH 1:800 BOSTER BM3876 Ki-67 1:200 BOSTER PB0065 Lamin B 1:1,000 Bioss bs-1840R β-action 1:800 Bioss bsm-33036M Open in a separate window caption a8 Primary antibodies with indicated concentration for WB, IHC, IF Cell proliferation, migration, and Matrigel invasion assay We used the Costar 24 well insert system (Life Technologies, corning, USA) to perform cell migration assay.

Techniques: In Vivo, Western Blot, Expressing